共培養(yǎng)體系中小鼠淋巴細胞RNA編輯酶ADAR1的表達及ADAR1在小鼠細胞免疫應答中的作用研究
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本文選題:共培養(yǎng) + 半定量PCR; 參考:《第四軍醫(yī)大學》2006年碩士論文
【摘要】:RNA編輯是近年來發(fā)現(xiàn)的一種遺傳信息修飾機制,它通過對前RNA特異位點的修飾,使腺嘌呤核苷轉化為次黃嘌呤核苷(Adenosine-to-inosine,A-to-Ⅰ),從而在轉錄后水平對目的基因進行修飾。接受編輯后的RNA翻譯產(chǎn)物可能會比基因組編碼的基因產(chǎn)物具有多樣性,除了傳統(tǒng)的蛋白翻譯途徑外,RNA編輯可以產(chǎn)生蛋白的同功異構體,從而在蛋白質多樣性的形成中發(fā)揮重要的作用。越來越多的研究發(fā)現(xiàn),在哺乳動物體內,許多與維持生命活動有關的重要功能蛋白質都受到A-to-ⅠRNA編輯的調控,使其結構和功能以及信號傳導通路都受到影響。目前發(fā)現(xiàn)A-to-ⅠRNA編輯酶的底物多局限于中樞系統(tǒng),而對Ⅰ-mRNA含量豐富的眾多外周組織中的A-to-ⅠRNA編輯情況的研究相對較薄弱。在哺乳類已經(jīng)克隆到四種RNA編輯酶(ADARs),即ADAR1、ADAR2、ADAR3、ADATs,它們具有相似的催化結構域。本實驗通過建立共培養(yǎng)細胞體系,觀察小鼠淋巴細胞在異種抗原細胞的刺激下ADAR1的表達情況;并通過siRNA質粒下調小鼠淋巴細胞ADAR1的表達,,觀察ADAR1表達抑制后的淋巴細胞免疫功能的變化。本實驗首次系統(tǒng)的對ADAR1在不
[Abstract]:RNA editor is a genetic modification mechanism that has been discovered in recent years. It modifies the adenine nucleoside (Adenosine-to-inosine, A-to- I) by modifying the special site of the former RNA, and modifies the target gene at the post transcriptional level. The edited RNA translation product may be more than the genome encoded gene. In addition to the traditional protein translation pathway, RNA editors can produce isoisomers of protein and play an important role in the formation of protein diversity. More and more studies have found that in mammals, many important functional proteins related to the maintenance of life activities are edited by A-to- I RNA. The structure, function and signal transduction pathway are affected. The substrate of the A-to- I RNA editing enzyme is found to be limited to the central system, and the study of A-to- I RNA in the many peripheral tissues with rich I -mRNA content is relatively weak. In mammalian, four RNA editing enzymes (ADARs), namely ADAR1, have been cloned. ADAR2, ADAR3, ADATs, they have a similar catalytic domain. In this experiment, a co culture cell system was established to observe the expression of ADAR1 in mouse lymphocytes under the stimulation of xenogeneic antigen cells, and the expression of ADAR1 in mouse lymphocytes was down regulated by siRNA plasmid, and the changes of lymphocyte immune function after the inhibition of ADAR1 expression were observed. The first system of this experiment on ADAR1 is not
【學位授予單位】:第四軍醫(yī)大學
【學位級別】:碩士
【學位授予年份】:2006
【分類號】:R392
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